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Marrow Stromal Cell-Based Cyclooxygenase 2 Ex Vivo Gene-Transfer Strategy Surprisingly Lacks Bone-Regeneration Effects and Suppresses the Bone-Regeneration Action of Bone Morphogenetic Protein 4 in a Mouse Critical-Sized Calvarial Defect Model

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Abstract

This study evaluated whether the murine leukemia virus (MLV)–based cyclooxygenase-2 (Cox-2) ex vivo gene-transfer strategy promotes healing of calvarial defects and/or synergistically enhances bone morphogenetic protein (BMP) 4–mediated bone regeneration. Gelatin scaffolds impregnated with mouse marrow stromal cells (MSCs) transduced with MLV-expressing BMP4, Cox-2, or a control gene were implanted into mouse calvarial defects. Bone regeneration was assessed by X-ray, dual-energy X-ray absorptiometry, and histology. In vitro, Cox-2 or prostanglandin E2 enhanced synergistically the osteoblastic differentiation action of BMP4 in mouse MSCs. In vivo, implantation of BMP4-expressing MSCs yielded massive bone regeneration in calvarial defects after 2 weeks, but the Cox-2 strategy surprisingly did not promote bone regeneration even after 4 weeks. Staining for alkaline phosphatase (ALP)–expressing osteoblasts was strong throughout the defect of animals receiving BMP2/4-expressing cells, but defects receiving Cox-2-expressing cells displayed weak ALP staining along the edge of original intact bone, indicating that the Cox-2 strategy lacked bone-regeneration effects. The Cox-2 strategy not only lacked bone-regeneration effects but also suppressed the BMP4-induced bone regeneration. In vitro coculture of Cox-2-expressing MSCs with BMP4-expressing MSCs in gelatin scaffolds reduced BMP4 mRNA transcript levels, suggesting that Cox-2 may promote BMP4 gene silencing in BMP4-expressing cells, which may play a role in the suppressive action of Cox-2 on BMP4-mediated bone formation. In summary, the Cox-2 ex vivo gene-transfer strategy not only lacks bone-regeneration effects but also suppresses the bone-regeneration action of BMP4 in healing of calvarial defects.

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Acknowledgements

This work was supported by a special appropriation to the Jerry L. Pettis Memorial VA Medical Center, Musculoskeletal Disease Center, and by an Assistance Award from the United States Army (DAMD-17-03-2-0021). The United States Army Medical Research Acquisition Activity (820 Chandler Street, Fort Detrick, MD 21702-5014) is the awarding and administering acquisition office. The information contained in this publication does not necessarily reflect the position or policy of the government, and no official endorsement should be inferred. All work was performed in facilities provided by the Department of Veterans Affairs.

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Authors and Affiliations

  1. Musculoskeletal Disease Center (151), Jerry L. Pettis Memorial VA Medical Center, 11201 Benton Street, Loma Linda, CA, 92357, USA

    K.-H. William Lau, Reinhard Gysin, Shin-Tai Chen, Jon E. Wergedal & Subburaman Mohan

  2. Department of Medicine, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA

    K.-H. William Lau, Reinhard Gysin, Shin-Tai Chen, Jon E. Wergedal, David J. Baylink & Subburaman Mohan

  3. Department of Biochemistry, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA

    K.-H. William Lau, Shin-Tai Chen, Jon E. Wergedal & Subburaman Mohan

  4. Department of Physiology, Loma Linda University School of Medicine, Loma Linda, CA, 92350, USA

    Subburaman Mohan

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  1. K.-H. William Lau
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Correspondence to K.-H. William Lau.

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Lau, KH.W., Gysin, R., Chen, ST. et al. Marrow Stromal Cell-Based Cyclooxygenase 2 Ex Vivo Gene-Transfer Strategy Surprisingly Lacks Bone-Regeneration Effects and Suppresses the Bone-Regeneration Action of Bone Morphogenetic Protein 4 in a Mouse Critical-Sized Calvarial Defect Model. Calcif Tissue Int 85, 356–367 (2009). https://doi.org/10.1007/s00223-009-9282-2

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  • DOI: https://doi.org/10.1007/s00223-009-9282-2

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